Targeted metabolomic analysis of phenylpropanoid metabolites in Astragalus membranaceus var. mongholicus in response to Fusarium solani infection / 药学学报

Root rot severely restricts the sustainable development of Astragalus membranaceus var. mongholicus (AMM) industry. Resistance breeding is an economical and environmentally safe way to manage the disease and its key lies in the obtaining of resistance indicators. This study aimed to quickly and accurately screen the resistance-related (RR) metabolites so as to provide reference for the screening of indicators of AMM breeding for resistance. LC-MS-based targeted metabolomics and real-time quantitative PCR technology were employed, in combination with multivariate statistical analysis, in analyzing the dynamic changes of phenylpropanoid metabolites in AMM in response to root rot pathogen Fusarium solani (FS) infection and identifying the differential metabolites. The LC-MS method established showed high sensitivity; each metabolite had a good linear relationship (R2 ≥ 0.968 9) in the corresponding linear range of the respective standard curve; the recoveries and the relative standard deviations (RSDs) (n = 6) ranged from 70% to 107% and from 1.2% to 9.9%, respectively. Obvious disturbances were observed in the changes of the targeted metabolites in AMM infected by FS. These metabolites, compared with the mock-inoculated (CK) group, showed different up or down regulation with time series. Calycosin-7-O-β-D-glucoside, ononin, calycosin and formononetin were identified as differential metabolites, and they all belong to flavonoids. The first three compounds were significantly negatively correlated (r ≤ -0.97, P < 0.05) with the content of FS in the root of AMM. As potential RR metabolites, they are helpful in obtaining promising resistance indicators for AMM against FS infection.

Caracterización genética de bacterias endofíticas de arroz (Oryza sativa L.) con actividad antimicrobiana contra Burkholderia glumae / Genetic characterization of rice endophytic bacteria (Oryza sativa L.) with antimicrobial activity against Burkholderia glumae

Resumen La demanda de xilanasas fúngicas en los procesos biotecnológicos industriales muestra un claro aumento en todo el mundo, por lo que hay un interés en ajustar las condicionesde producción de xilanasas microbianas. En este estudio se optimizó la capacidad del hongoFusarium solani para producir xilanasas extracelulares con escasa actividad celulolítica medi-ante el dise˜no de Box-Wilson. Se determinaron las mejores condiciones de cultivo para obteneruna preparación enzimática cruda con una actividad xilanolítica significativa y poca actividad celulolítica. En la mayoría de los tratamientos, la actividad xilanolítica fue mayor que laactividad celulolítica. Se observó un efecto negativo sobre la producción de endoxilanasas, xilosidasas y endocelulasas con el aumento de la concentración de xilano. El aumento del tiempode incubación afectó adversamente la producción de endocelulasas y xilosidasas. De acuerdocon el modelo matemático y las pruebas experimentales, es posible producir endoxilanasas conuna actividad endocelulasa mínima aumentando el tiempo de incubación y la concentración desulfato de amonio. Las condiciones de cultivo óptimas para producir una mayor cantidad deendoxilanasas (10,65 U/mg) y mínima cantidad de endocelulasas fueron 2,5% (p/v) de xilano y5, 2 y 0,4 g/l de extracto de levadura, sulfato de amonio y urea, respectivamente, con 120 hde incubación.

Abstract The aim of the present study was to isolate, select and characterize endophytic bacteria in rice inhibiting Burkholderia glumae THT as well as to characterize the genetic diversity and virulence factors in strains of B. glumae and Burkholderia gladioli of rice. Rice plants were collected in 4 departments from the northern region of Peru, isolating endophytic bacteria, aftertissue sterilization, at 30°C (48 h) in Trypticase SoyAgar (TSA), evaluating the antimicrobial activity against B. glumae THT, production of siderophores, resistance of toxoflavine and partial sequencing of the 16S rRNA gene. Furthermore, B. glumae and B. gladioli were isola-ted in selective medium (pH 4.5) at 41 °C/72h. Molecular identification was performed using BOX-PCR and sequencing of the 16S rRNA gene, in addition to the production of extracellular enzymes, motility tests and sensitivity/resistance to bactericides. One hundred and eighty nine (189) endophytic bacteria were isolated, and only 9 strains showed antimicrobial activity against B. glumae THT, highlighting Burkholderia vietnamiensis TUR04-01, B. vietnamiensis TUR04-03 and Bacillus aryabhattai AMH12-02. The strains produced siderophores and at least 55.5% were resistant to toxoflavin. Additionally, 17 strains were grouped into 9 BOX-PCR profiles, where 16 had similarity with B. glumae LMG2196T (100%) and 1 with B. gladioli NBRC 13700T (99.86%). High diversity was found according to geographical origin and virulence factors. In conclusion, strains of the genus Bacillus and Burkholderia are potential biocontrol agents against B. glumae.

Optimization of production of xylanases with low cellulases in Fusarium solani by means of a solid state fermentation using statistical experimental design / Optimización de la producción de xilanasas con celulasas bajas en Fusarium solani mediante una fermentación del estado sólido utilizando diseño experimental estadístico

Resumen La demanda de xilanasas fúngicas en los procesos biotecnológicos industriales muestra un claro aumento en todo el mundo, por lo que hay un interés en ajustar las condiciones de producción de xilanasas microbianas. En este estudio se optimizó la capacidad del hongo Fusarium solani para producir xilanasas extracelulares con escasa actividad celulolítica mediante el diseño de Box-Wilson. Se determinaron las mejores condiciones de cultivo para obtener una preparación enzimática cruda con una actividad xilanolítica significativa y poca actividad celulolítica. En la mayoría de los tratamientos, la actividad xilanolítica fue mayor que la actividad celulolítica. Se observó un efecto negativo sobre la producción de endoxilanasas, p-xilosidasasy endocelulasascon el aumento de la concentración dexilano. El aumento del tiempo de incubación afectó adversamente la producción de endocelulasas y p-xilosidasas. De acuerdo con el modelo matemático y las pruebas experimentales, es posible producir endoxilanasas con una actividad endocelulasa mínima aumentando el tiempo de incubación y la concentración de sulfato de amonio. Las condiciones de cultivo óptimas para producir una mayor cantidad de endoxilanasas (10,65 U/mg) y mínima cantidad de endocelulasas fueron 2,5% (p/v) de xilano y 5, 2 y 0,4 g/l de extracto de levadura, sulfato de amonio y urea, respectivamente, con 120 h de incubación.

Resumen La demanda de xilanasas fúngicas en los procesos biotecnológicos industriales muestra un claro aumento en todo el mundo, por lo que hay un interés en ajustar las condicionesde producción de xilanasas microbianas. En este estudio se optimizó la capacidad del hongo Fusarium solani para producir xilanasas extracelulares con escasa actividad celulolítica medi-ante el dise˜no de Box-Wilson. Se determinaron las mejores condiciones de cultivo para obteneruna preparación enzimática cruda con una actividad xilanolítica significativa y poca actividad celulolítica. En la mayoría de los tratamientos, la actividad xilanolítica fue mayor que laactividad celulolítica. Se observó un efecto negativo sobre la producción de endoxilanasas, xylanolytic activity and little cellulolytic activity. In most treatments, the xylanolytic activity was higher than the cellulolytic activity. A negative effect on the production of endoxylanases, p-xylosidases and endocellulases was observed with the increasing of xylan concentration. Increasing the incubation time adversely affected the production of endocellulases and p-xylosidases. According to the mathematical model and experimental tests, it is possible to produce endoxylanases with minimal endocellulase activity increasing incubation time and the concentration of ammonium sulfate. The optimal culture conditions to produce a greater amount of endoxylanases (10.65 U/mg) and low endocellulases from F. solani were: 2.5% (w/v) xylan, 5.0, 2.0 and 0.4g/l, of yeast extract, ammonium sulfate and urea, respectively, with 120 h of incubation.

Phytochemistry and antifungal potential of Datura inoxia Mill. on soil phytopathogen control / Fitoquímica e potencial antifúngico de Datura inoxia Mill. no controle de fitopatógenos do solo

The application of chemical pesticides for the control of fungal diseases results in impacts on the environment and human health. The use of vegetal extracts with antifungal properties for the proper management of crops becomes a viable alternative, mainly for organic and family farming. The objective of this study was to carry out the phytochemical evaluation of Datura inoxia, evaluating its antifungal potential against the mycelial growth of Fusarium solani and Sclerotinia sclerotiorum. The extracts, aqueous and ethanolic, obtained from the leaves of the plant collected in areas of the municipality of Campo Grande, Mato Grosso do Sul, were submitted to phytochemical prospecting and quantification of flavonoids and total phenols. It was evaluated its antifungal activity at concentrations of 800, 1200, 1600, 2000, and 2400 µg 100 mL-1. Each concentration was separately incorporated into BDA agar, poured into Petri dishes, and inoculated with the mycelial disc of the fungus. The diameter of the colonies were measured daily. Two solutions were prepared as control, one containing the solvent added to PDA medium (ethanol solution), and another with only PDA medium (without D. inoxia extract, control). In both extracts were found the same diversity of secondary metabolites (nine classes). The ethanolic extract, a solvent of lower polarity than water, was more efficient in the extraction of these constituents. Alkaloids and phenolic compounds were the most frequent compounds (100%). In relation to antifungal activity, the ethanolic extract provided 100% inhibition of mycelial growth ofSclerotinia sclerotitorum in all concentrations, relative to the control. On the other hand, the growth ofFusarium solani was only negatively affected at the highest concentrations of 800 and 1200 µmL-1 100 mL-1. The antifungal potential of Datura inoxia was probably related to the abundance of alkaloids and phenolic compounds in its chemical constitution that negatively effects the development of the vegetative mycelium.

A aplicação de defensivos químicos para o controle de doenças fúngicas tem por consequência impactos sobre o ambiente e a saúde humana. Desta forma, a utilização de extratos vegetais com propriedades antifúngicas associado ao manejo adequado de culturas, torna-se uma proposta viável de controle alternativo, principalmente na agricultura orgânica e familiar. Neste sentido, objetivou-se realizar a avaliação fitoquímica das folhas de Datura inoxia, avaliando seu potencial antifúngico frente ao crescimento micelial de Fusarium solani e Sclerotinia sclerotiorum. Os extratos, aquoso e etanólico, obtidos das folhas da planta coletadas em áreas do município de Campo Grande, Mato Grosso do Sul, foram submetidos à prospecção fitoquímica e quantificação flavonoides e fenóis totais, avaliando-se sua atividade antifúngica em concentrações de 800, 1200, 1600, 2000 e 2400 µg 100 mL-1. Cada concentração foi incorporada, separadamente, em ágar BDA, vertida em placas de petri, seguida da colocação do disco de micélio do fungo, com diâmetro das colônias sendo medido diariamente. Utilizou-se como controle negativo, ágar sem extrato e ágar com solução etanólica. Nos dois extratos ocorreu a mesma diversidade de metabólitos secundários (nove classes); porém o extrato etanólico, um solvente de menor polaridade que a água, foi mais eficiente na extração destes constituintes, com destaque aos alcaloides e compostos fenólicos com maior frequência (100%). Em relação a atividade antifúngica, o extrato etanólico proporcionou inibição de 100% do crescimento micelial de Sclerotinia sclerotitorum, em todas as concentrações, em relação a testemunha. Por outro lado, o crescimento de Fusarium solani foi afetado negativamente apenas nas maiores concentrações, 800 e 1200 µmL-1 100 mL-1.O potencial antifúngico da planta provavelmente está relacionado a sua constituição química, com abundância de alcaloides e compostos fenólicos, afetando negativamente o desenvolvimento do micélio vegetativo.

Effects of combination of phenazine-1-carboxylic acid from Streptomyces kebangsaanensis with amphotericin B on clinical Fusarium solani species

Aim@#A novel endophyte, Streptomyces kebangsaanensis was isolated from the stem of a Malaysian ethnomedicinal plant, Portulaca oleracea in 2013. Studies on S. kebangsaanensis crude extract showed that it had antifungal activities and further work led to isolation of a novel compound, phenazine-1-carboxylic acid (PCA). This study investigated the combinatorial effect of PCA isolated from S. kebangsaanensis with amphotericin B on the growth of four clinical Fusarium solani isolates. @*Methodology and results@#Disk diffusion assay showed that the crude extract of S. kebangsaaneesis inhibited growth of all four F. solani isolates. Whereas, the compound PCA from this extract inhibited two of the tested F. solani isolates, UZ541/12, and UZ667/13 at minimum inhibitory concentration of 18.00 µg/mL Combinations of this compound with amphotericin B, reduced the minimum inhibitory concentration of amphotericin B for these two isolates from 8 to 0.13 µg/mL and 4 to 0.03 µg/mL respectively. Analysis of fractional inhibitory concentration index showed that a borderline synergism is present between the compound and amphotericin B. @*Conclusion, significance and impact of the study@#These results indicate PCA may be useful in improving actions of available drugs against antimicrobial resistant microorganisms.

Real-time PCR detection method for Fusarium solani, pathogen of Panax notoginseng root rot / 中草药

Objective: In order to establish an accurate and rapid real-time fluorescence quantitative PCR method for the detection of Fusarium solani, the pathogenic fungus of Panax notoginseng root rot. Methods: Based on aminoadipate reductase Lys2 gene of F. solani, specific primers Fs-QF and Fs-QR were designed. The recombinant plasmid standard was prepared and the SYBR Green I fluorescence real-time quantitative PCR method and real-time fluorescent 1oop-mediated isothermal amplication (LAMP) system for detecting F. solani was established. Fifteen samples including P. notoginseng plants with typical symptoms of root rot and black spot as well as soil of P. notoginseng planting area were collected. The total DNA of these samples were extracted as templates, and then detected by the real-time fluorescence quantitative PCR method and 1oop-mediated isothermal amplification established in this study. Results: The real-time fluorescent quantitative PCR method and 1oop-mediated isothermal amplification technique established in this study had high specificity. P. notoginseng plants infected with F. solani can be detected quickly. In addition, the method has high sensitivity and the concentration of detection template can be as low as 0.2 pg/μL. Conclusion: The method established in this study can be used to reveal the dynamic changes of F. solani concentration in the complex P. notoginseng planting soil and diseased plants. Furthermore, it can provide technical supports for the soil treatment, early diagnosis and dynamic monitoring of root rot, and rapid molecular detection of P. notoginseng seeds and seedlings.

<i>In vitro</i> interactions between antifungals and Cyclosporin A against Fusarium solani / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the<i> in vitro</i> interaction between antifungals and tacrolimus acting alone or in combination against Fusarium solani.<p>METHODS: According to Clinical and Laboratory Standards Institute(CLSI)M27-Ed4 and M38-A3, 22 strains of Fusarium solani were used to perform drug sensitivity tests with chessboard microdilution method by cyclosporin A combined with 4 kinds of antifungal drugs <i>in vitro</i>.<p>RESULTS: The MIC ranges of natamycin, voriconazole, amphotericin B and fluconazole against 22 strains of Fusarium solani were 2-8, 1-8, 1-8 and 8-512μg/mL respectively. When combined with tacrolimus <i>in vitro</i>, the synergistic effects of fluconazole and Amphotericin B were observed in 64% and 41% strains respectively. There were no antagonistic effects observed in all combined drug tests. With the combination, the sensitivity of Fusarium to amphotericin B was significantly increased from 4.5% to 68.2%(<i>P</i><0.001).<p>CONCLUSION: Fusarium solani is sensitive to natamycin <i>in vitro</i> and is partially sensitive to voriconazole. When combined with cyclosporine A, it can produce synergistic effects with fluconazole and amphotericin B, and significantly increase the sensitivity of Fusarium solani to amphotericin B drugs.

Optimization of camptothecin production and biomass yield from endophytic fungus Fusarium solani strain ATLOY-8

Fusarium solani strain ATLOY-8, a fungus that produces camptothecin (CPT) was isolated from Chonemorphafragrans (Moon) Alston. The process of optimization of the camptothecin yield was optimized by means of thetraditional methods and analytical tools. Using the Box–Behnken design (BBD) matrix at N = 17, three independentvariables were optimized designated using one factor at a time (OFAT) method, for improved CPT and biomassproduction. The BBD exhibited 1.4- and 1.2-fold increase of CPT production and biomass yield, respectively,compared to OFAT approach in the basal medium containing absolute ethanol (5%), glucose (1%), and precursors (acombination of tryptophan, geraniol, and tryptamine; 0.03%). Analysis of variance showed the elevated coefficient ofdetermination (R2) of 0.9751 and 0.9980, respectively, for CPT and biomass yielding at a significant level (p > 0.05).Three dimensional graphs revealed dependent synergy between two variables to improve the CPT and biomass yield

Fusarium solani keratitis: role of antifungal susceptibility testing and identification to the species level for proper management

ABSTRACT We report a patient with fungal keratitis caused by a multiresistant Fusarium solani in a tertiary care hospital located in southern Brazil. A 55-year-old man with a history of ocular trauma presented with keratitis in left eye. The patient has a complicated clinical course and failed to respond to local and systemic antifungal treatment, and required eye enucleation. Despite multiple topical, intraocular and systemic antifungal treatments, hyphal infiltration persisted in the corneal transplant causing continuous recurrences. The cultures of corneal biopsy scrapings were positive for Fusarium spp. The organism was identified to species level by multi-locus sequencing for translation elongation factor 1 alpha (EF-1α), and RNA polymerase II subunit (RPB2). In vitro antifungal susceptibility testing of the isolate by the broth microdilution method, according to CLSI M38-A2, disclosed susceptibility to natamycin and resistance to amphotericin B, voriconazole, itraconazole and fluconazole. Considering previous unsuccessful antifungal treatments due to multiple drug resistance, the eye was enucleated. Our case report illustrates that management of fungal keratitis remains a therapeutic challenge. Optimal treatment for F. solani infection has not yet been established and should include susceptibility testing for different antifungal agents.

Isolation and structural elucidation of antifungal compounds from Curcuma amada

Objective: To isolate and identify the antifungal compounds from Curcuma amada. Methods: The antifungal activity was measured by the diameter of colonies grown on Petri dish, microscopic observation, and CLSI microdilution methods. The antifungal compounds were isolated through bioactivity guided purification by using silica gel and high-performance liquid chromatography. Structural identification of the antifungal compounds was conducted using

Isolation and structural elucidation of antifungal compounds from Curcuma amada

Objective: To isolate and identify the antifungal compounds from Curcuma amada. Methods: The antifungal activity was measured by the diameter of colonies grown on Petri dish, microscopic observation, and CLSI microdilution methods. The antifungal compounds were isolated through bioactivity guided purification by using silica gel and high-performance liquid chromatography. Structural identification of the antifungal compounds was conducted using 1H NMR, 13C NMR, and liquid chromatography-tandem mass spectrometry. Results: The purified antifungal compounds were zederone and furanodienone. These two compounds showed dose-dependent antifungal activity against Fusarium solani sensu lato. The concentration required for 50% growth inhibition (IC50) of FSSL ranged from 115 to 129 μM and 82 to 91 μM for zederone and furanodienone, respectively. Conclusion: This study suggested that the isolated compounds from Curcuma amada could be promising natural antifungal agents to control the diseases caused by Fusarium solani sensu lato.

Inhibition effect of biocontrol bacteria NJ13 and its mixture with chemical fungicides against ginseng root rot caused by Fusarium solani / 中国中药杂志

This study was aimed to clarify the toxicity indoor and inhibition effect of biocontrol strain NJ13 and its mixture with chemical fungicides against Fusarium solani causing ginseng root rot. The method of mycelial growth rate and Sun Yunpei method were used to determine the indoor toxicity and co-toxicity coefficient of strain NJ13 and their mixture with chemical pesticides against F. solani. The dual culture assay method,mixed culture method and microscopic observation were used to determine the sporulation and germination of spores and mycelial growth and morphological change of hyphae of F. solani treated by strain NJ13. The results of toxicity indoor showed that strain NJ13 had the best inhibitory effect on pathogen,and its EC_(50) value was 0. 071 mg·L~(-1). It was all synergistic for antifungal effect that strain NJ13 was mixed with propiconazole and difenoconazole respectively with a range from 1 ∶4 to 4 ∶1( volume ratio). Both of optimal ratios were 1 ∶1,and the co-toxicity coefficients were 848. 70 and 859. 73,respectively. The strain NJ13 could inhibit the sporulation,germination and mycelial growth of F. solani. The biocontrol strain NJ13 had an inhibition effect on F. solani,and the optimal antifungal ratio of strain NJ13 mixed with propiconazole and difenoconazole was obtained.

Essential oils of Myrcia lundiana Kiaersk and their major compounds show differentiated activities against three phytopathogenic fungi / Óleos essenciais de Myrcia lundiana Kiaersk e seus compostos majoritários apresentam atividades diferenciadas sobre três fungos fitopatogênicos

The present study evaluated the antifungal activity of the essential oils of chemotypes of Myrcia lundiana and their major compounds on the fungi Fusarium pallidoroseum, Fusarium solani, and Colletotrichum musae. The essential oils were obtained by hydrodistillation and analyzed by GCMS/FID. For the evaluation of the antifungal activity, the essential oils and the major compounds were tested at the concentration of 0.1 mL/L until the fungicidal effect was detected. The major compounds detected in the essential oil were 1,8-cineole, isopulegol, and citral. The chemotypes (MLU-005 and MLU-019) provided 100% mycelial growth inhibition for the fungus F. pallidoroseum from the concentration of 1.1 mL/L (minimum inhibition concentration - MIC). For chemotype (MLU-022), the minimum fungicidal concentration (MFC) was 0.3 mL/L. For F. solani, the essential oils of the chemotypes (MLU-005 and MLU-019) presented MIC at concentrations of 7.0 and 5.0 mL/L, respectively. The essential oil of the chemotype (MLU-022) presented MFC of 0.6 mL/L. Different MIC was observed for the three studied chemotypes for the fungus C. musae, ranging between 0.4 mL/L, for the chemotype (MLU-005); 0.5 mL/L, for the chemotype (MLU-022); and 0.7 mL/L, for the chemotype (MLU-019). The best MFC was observed for the chemotype (MLU-005) (0.5 mL/L). The major compounds tested separately presented better MIC values when compared with their chemotypes, except for the compound 1,8-cineole, which presented lower mycelial growth inhibition for the three fungi tested, suggesting that the chemical profile or the presence of some other compound of the essential oil may inhibit the growth of the three fungi studied. The compound isopulegol provided lower MFC for the fungus C. musae (0.4517 mL/L) when compared with the fungi F. pallidoroseum and F. solani, (MFC of 0.4927 mL/L). The compound citral provided a lower MFC on the fungus C. musae (0.1668 mL/L) in relation to the other fungi tested. The essential oils of the chemotypes of M. lundiana and their major compounds showed potential to control the studied phytopathogens and can be an alternative for agriculture for presenting an inhibitory and fungicidal effect against these organisms at lower concentrations.

O presente trabalho avaliou a atividade antifúngica de óleos essenciais de quimiotipos de Myrcia lundiana dos seus compostos majoritários sobre os fungos Fusarium pallidoroseum, Fusarium solani e Colletotrichum musae. Os óleos essenciais foram obtidos por hidrodestilação e analisados por CGEM/DIC. Para avaliação da atividade antifúngica, foram testados os óleos essenciais e os compostos majoritários na concentração de 0,1 mL/L até encontrar o efeito fungicida. Os principais compostos presentes no óleo essencial foram 1,8-cineol, isopulegol e citral. Os quimiotipos (MLU-005 e MLU-019) proporcionaram 100% de inibição do crescimento micelial para o fungo F. pallidoroseum a partir da concentração de 1,1 mL/L (Concentração Inibitória Mínima ­ CIM). Para o quimiotipo (MLU-022), a melhor concentração fungicida mínima (CFM) foi de 0,3 mL/L. Para F. solani, os óleos essenciais dos quimiotipos (MLU-005 e MLU-019) apresentaram CIM nas concentrações de 7,0 e 5,0 mL/L, respectivamente. O óleo essencial do quimiotipo (MLU-022) apresentou CFM de 0,6 mL/L. Observou-se CIM diferenciado para os três quimiotipos estudados para o fungo C. musae, variando entre 0,4 mL/L, para o quimiotipo (MLU-005); 0,5 mL/L, para o quimiotipo (MLU-022); e 0,7 mL/L, para o quimiotipo (MLU-019). O quimiotipo MLU-005 apresentou o melhor CFM, 0,5 mL/L. Os compostos majoritários testados separadamente apresentaram melhores valores de CIM frente aos seus quimiotipos, exceto o composto 1,8-cineol, que apresentou menor inibição do crescimento micelial para os três fungos testados, sugerindo que o perfil químico ou a presença de algum outro composto no óleo essencial pode estar atuando na inibição do crescimento dos três fungos estudados. O composto isopulegol proporcionou menor CFM para o fungo C. musae (0,4517 mL/L) em relação aos fungos F. pallidoroseum e F. solani, para os quais apresentou CFM de 0,4927 mL/L. O composto citral proporcionou um menor CFM sobre o fungo C. musae (0,1668 mL/L), em relação aos demais fungos testados. Os óleos essenciais de quimiotipos de M. lundiana e seus compostos majoritários apresentaram potencial para o controle dos fitopatógenos estudados, podendo ser considerados como uma alternativa para a agricultura, uma vez que em concentrações mais baixas apresentaram efeito inibitório e fungicida frente a estes organismos.

Activity of essential oils of Lippia alba chemotypes and their major monoterpenes against phytopathogenic fungi / Atividade dos óleos essenciais de quimiotipos de Lippia alba e seus monoterpenos majoritários sob fungos fitopatogênicos

This work aimed to evaluate the in vitro antifungal activity of the essential oils of L. alba belonging to the carvone chemotype (LA-13 and LA-57) and the citral chemotype (LA-10, LA-29, and LA-44); the carvone enantiomers (R)-(-)-carvone and (S)-(+)-carvone; and citral on phytopathogenic fungi Lasiodiplodia theobromae (LT), Fusarium pallidoroseum (FP) and Fusarium solani (FS). Concentrations of 0.01; 0.05; 0.1; 0.2; 0.3; 0.5 and 1.0 mL/100 mL were tested, and the percentage of mycelial growth inhibition (MGI) was calculated after 96h in relation to the control. Minimal Inhibitory Concentrations (MIC) and Minimal Fungicide Concentrations (MFC) were obtained for essential oils and compounds. From the concentration of 0.2 mL/100 mL, all the accessions and carvone enantiomers were effective against the fungus LT, except the accession LA-44, for which the maximum inhibition occurred from the concentration of 0.3 mL/100 mL. Citral was the most effective compound against LT, with 100% of MGI from the concentration of 0.05 mL /100 mL. All accessions and enantiomers caused 100% of MGI against FP fungus from the concentration of 0.2 mL/100 mL. Once again, citral stood out by providing the same result as the other treatments from the concentration of 0.1 mL/100 mL. Considering the fungus FS, carvone enantiomers and citral caused 100% of MGI from the concentration of 0.1 mL/100 mL while all accessions caused 100% of MGI from the concentration of 0.2 mL/100 mL. Citral and carvone enantiomers presented the lowest MIC values (0.1 mL/100 mL) against FS fungus. The MIC of citral for LT and FP were not determined at the concentrations tested. (R)-(-)-carvone enantiomer presented the lowest MIC (0.1 mL/100 mL) for the LT fungus. Most of the other accessions presented MIC of 0.2 mL/100 mL for the three fungi. In relation to the minimum fungicidal concentration (MFC), citral stood out with values from 0.05 mL/100 mL (LT). Citral and carvone presented the same MFC for FS (0.2 mL / 100 mL). The other accessions showed MFC values from 0.3 mL/100 mL for the three fungi. Essential oils of L. alba accessions, carvone enantiomers, and citral were efficient in phytopathogen control and could be considered as an alternative to fungicides for presenting inhibitory and fungicidal effect against these microorganisms at low concentrations.

O objetivo do trabalho foi avaliar a atividade antifúngica in vitro de óleos essenciais de Lippia alba pertencentes ao quimiotipo carvona (LA-13 e LA-57) e ao quimiotipo citral (LA-10, LA-29 e LA-44); dos enantiômeros da carvona: (R)-(-)-carvona e (S)-(+)-carvona; e do citral sobre os fungos fitopatogênicos Lasiodiplodia theobromae (LT), Fusarium pallidoroseum (FP) e Fusarium solani (FS). Foram testadas as concentrações 0,01; 0,05; 0,1; 0,2; 0,3; 0,5 e 1,0 mL/100 mL, e, após 96h de incubação, a porcentagem de inibição do crescimento micelial (ICM) foi calculada em relação ao controle. Foram determinadas as Concentrações Inibitórias Mínimas (CIM) e Fungicidas Mínimas (CFM) para os óleos essenciais e compostos. A partir da concentração de 0,2 mL/100 mL todos os acessos e os enantiômeros da carvona foram efetivos contra LT, exceto o acesso LA-44, que proporcionou máxima inibição a partir da concentração de 0,3 mL/100 mL. O monoterpeno citral foi o mais efetivo contra LT, pois a partir da concentração de 0,05 mL /100 mL, 100% de ICM foi observada. Todos os acessos e enantiômeros da carvona causaram 100% de ICM contra o fungo FP, a partir da concentração de 0,2 mL/100 mL. Novamente, o composto citral de destacou por causar máxima ICM a partir da concentração de 0,1 mL/100 mL. Contra o fungo FS, os enantiômeros da carvona e o citral causaram 100% de ICM a partir da concentração de 0,1 mL/100 mL, enquanto os acessos proporcionaram mesmos resultados a partir da concentração de 0,2 mL/100 mL. O citral e os enantiômeros da carvona apresentaram os menores valores de CIM (0,1 mL/100 mL) frente ao FS. Não foi possível determinar a CIM do citral para LT e FP nas concentrações testadas. O enantiômero (R)-(-)-carvone apresentou a menor CIM (0,1 mL/100 mL) para o fungo LT. Os acessos apresentaram CIM a partir de 0,2 mL/100 mL para os três fungos. Em relação à concentração fungicida mínima (CFM), o citral se destacou com a menor CFM (0,05 mL/100 mL) para LT. Citral e carvonas apresentaram a mesma CFM para FS (0,2 mL / 100 mL). Os acessos apresentaram CFM a partir de 0,3 mL/100 mL para os três fungos. Os óleos essenciais dos acessos de L. alba, e os monoterpenos carvona e o citral foram eficientes no controle dos fungos fitopatogênicos e são considerados como uma alternativa em relação aos fungicidas sintéticos por apresentarem efeitos inibitórios e fungicidas contra esses microorganismos quando utilizados em baixas concentrações.

Efecto de recubrimientos de almidón modificado de yuca, proteina aislada de soya y aceite esencial de orégano aplicados a la papaya / Effect of modified yuca starch coatings, isolated protein of soya and oregano essential oil applied to the papaya

RESUMEN La papaya tiene elevada demanda en el mundo; sin embargo, la producción y la exportación se afectan por microorganismos fitopatógenos y pérdidas pos cosecha, cercanas al 25-40% en la cadena de suministro, sobre todo, en transporte y en almacenamiento, debido a un deficiente manejo. La aplicación de recubrimientos comestibles permite mejorar el brillo y la textura de la corteza, reducir el deterioro de la calidad fisicoquímica y organoléptica, la pérdida de peso por deshidratación y el intercambio de gases. Por lo anterior, durante trece días, en papayas recubiertas con almidón de yuca modificado variedad SM 707-17 (4%), proteína aislada de soya (2%) y aceite esencial de orégano (250ppm y 500ppm), se evaluó el efecto sobre pérdida de peso, de color, de firmeza, de sólidos solubles, de respiración, de pH, de acidez titulable y de crecimiento del Fusarium spp., a condiciones ambientales, mediante un diseño completamente al azar con dos factores, tiempo y tratamientos. El experimento, se hizo por triplicado y los datos fueron sometidos a un análisis estadístico, utilizando el programa SPSS V.23. Los resultados indicaron que la utilización de almidón, por sí solo o combinado con aceite esencial de orégano (250ppm) y proteína aislada, lograron reducir la pérdida de peso, controlar la respiración al reducir el porcentaje de CO2 y retardar el desarrollo del color amarillo y rojo en las papayas frente al control, además de retrasar el crecimiento del hongo Fusarium solani; también, se encontró que los recubrimientos no incidieron sobre características fisicoquímicas, como el pH, la acidez titulable, los sólidos solubles y la firmeza de los frutos.

SUMMARY The papaya has a high demand in the world, however, its production and export are affected by phytopathogenic microorganisms and post-harvest losses close to 25-40% in the supply chain, especially in transport and storage due to poor management. The application of edible coatings improves the gloss and texture of the bark, reduces the deterioration of the physicochemical and organoleptic quality, the weight loss due to dehydration, and the exchange of gases. Therefore, for thirteen days, papayas were coated with cassava starch modified SM 707-17 variety (4%), isolated soy protein (2%) and oregano essential oil (250ppm and 500ppm). The effect was evaluated on weight loss, color, firmness, soluble solids, respiration, pH, titulable acidity and growth of Fusarium spp., to environmental conditions. A completely random design with two factors, time and treatments, the experiment was done in triplicate, and the data were subjected to a statistical analysis using the SPSS V.23 program. The results indicated that the use of starch alone or combined with essential oil of oregano (250ppm) and isolated protein were able to reduce weight loss, control respiration (CO2 percent) and slow down the development of the yellow and red color in the papayas against the control in addition to delaying the growth of the fungus Fusarium spp. It was also found that the coatings did not affect physicochemical characteristics such as pH, titulable acidity, soluble solids, and firmness of the fruits.

Phytochemistry, antioxidant potential and antifungal of Byrsonima crassifolia on soil phytopathogen control / Fitoquímica, potencial antioxidante e antifúngico de Byrsonima crassifolia no controle de fitopatógenos de solo

Abstract The use of chemical defensives to control fungal diseases has by consequence to impact negatively over the environment and human health, this way, the use of plant extracts with antifungal properties along with proper cultural management makes viable an alternative plant production control, specially for familiar and organic cultures. The objective of this study was to perform phytochemical and antioxidant analysis of Byrsonima crassifolia (canjiqueira) barks and evaluate its antifungal potential over Fusarium solani and Sclerotinia sclerotiorum mycelial growth. The ethanol extract from plants collected in Pantanal, Mato Grosso do Sul, Brazil was submitted to phytochemical prospection, total phenol and flavonoids quantification and antioxidant activiy determination (DPPH). To evaluate antifungal activity concentrations of 800, 1200, 1600, 2000 and 2400 µg 100 mL-1 of ethanol extract were used. Which concentration was separately incorporated in agar (PDA) and shed in Petri dishes, followed by the fungi mycelial disc where the colonies diameter was measured daily. Negatives control with agar without extract and agar with an ethanol solution were used. The B. crassifolia ethanol extract presented inhibitory activity over the fungi studied where concentrations of 800 and 1600 µg 100 mL-1, inhibited 38% of the mycelial growth of F. solani; to S. sclerotiorum the best concentration was 2400 µg 100 mL1, reducing 37.5%. The antifungal bark extract potential of this specie is attributed to phenolic compounds and to triterpenes derivatives.

Resumo A aplicação de defensivos químicos para o controle de doenças fúngicas tem por consequência impactos sobre o ambiente e a saúde humana, dessa forma, a utilização de extratos vegetais com propriedades antifúngicas associado ao manejo adequado de culturas, torna-se uma proposta viável de controle alternativo, principalmente na agricultura orgânica e familiar. Neste sentido, objetivou-se neste trabalho realizar a análise fitoquímica e antioxidante das cascas de Byrsonima crassifolia (canjiqueira) e avaliar seu potencial antifúngico sobre o crescimento micelial de Fusarium solani e Sclerotinia sclerotiorum. O extrato etanólico das cascas da planta, coletadas no Pantanal do Rio Negro, em Mato Grosso do Sul, foi submetido à prospecção fitoquímica, quantificação de fenóis totais e flavonoides e determinação da atividade antioxidante (DPPH). Para a avaliação da atividade antifúngica foram utilizadas as concentrações de 800, 1200, 1600, 2000 e 2400 µg 100 mL-1 do extrato etanólico. Cada concentração foi incorporada, separadamente, em ágar BDA, e vertida em placas de petri, seguido do disco de micélio do fungo, onde o diâmetro das colônias foi medido diariamente. Utilizou-se como controle negativo, ágar sem extrato e ágar com solução etanólica. O extrato etanólico de B. crassifolia apresentou atividade inibitória sobre os fungos estudados, onde as concentrações de 800 e 1600 µg 100 mL-1, inibiram 38% do crescimento micelial de F. solani; para S. sclerotiorum, a melhor concentração foi de 2400 µg 100 mL1, com 37,5% de redução de crescimento. Atribui-se o potencial antifúngico do extrato da casca da espécie aos compostos fenólicos e derivados de triterpenos.

Phytochemistry, antioxidant potential and antifungal of Byrsonima crassifolia on soil phytopathogen control

Abstract The use of chemical defensives to control fungal diseases has by consequence to impact negatively over the environment and human health, this way, the use of plant extracts with antifungal properties along with proper cultural management makes viable an alternative plant production control, specially for familiar and organic cultures. The objective of this study was to perform phytochemical and antioxidant analysis of Byrsonima crassifolia (canjiqueira) barks and evaluate its antifungal potential over Fusarium solani and Sclerotinia sclerotiorum mycelial growth. The ethanol extract from plants collected in Pantanal, Mato Grosso do Sul, Brazil was submitted to phytochemical prospection, total phenol and flavonoids quantification and antioxidant activiy determination (DPPH). To evaluate antifungal activity concentrations of 800, 1200, 1600, 2000 and 2400 µg 100 mL-1 of ethanol extract were used. Which concentration was separately incorporated in agar (PDA) and shed in Petri dishes, followed by the fungi mycelial disc where the colonies diameter was measured daily. Negatives control with agar without extract and agar with an ethanol solution were used. The B. crassifolia ethanol extract presented inhibitory activity over the fungi studied where concentrations of 800 and 1600 µg 100 mL-1, inhibited 38% of the mycelial growth of F. solani; to S. sclerotiorum the best concentration was 2400 µg 100 mL1, reducing 37.5%. The antifungal bark extract potential of this specie is attributed to phenolic compounds and to triterpenes derivatives.

Resumo A aplicação de defensivos químicos para o controle de doenças fúngicas tem por consequência impactos sobre o ambiente e a saúde humana, dessa forma, a utilização de extratos vegetais com propriedades antifúngicas associado ao manejo adequado de culturas, torna-se uma proposta viável de controle alternativo, principalmente na agricultura orgânica e familiar. Neste sentido, objetivou-se neste trabalho realizar a análise fitoquímica e antioxidante das cascas de Byrsonima crassifolia (canjiqueira) e avaliar seu potencial antifúngico sobre o crescimento micelial de Fusarium solani e Sclerotinia sclerotiorum. O extrato etanólico das cascas da planta, coletadas no Pantanal do Rio Negro, em Mato Grosso do Sul, foi submetido à prospecção fitoquímica, quantificação de fenóis totais e flavonoides e determinação da atividade antioxidante (DPPH). Para a avaliação da atividade antifúngica foram utilizadas as concentrações de 800, 1200, 1600, 2000 e 2400 µg 100 mL-1 do extrato etanólico. Cada concentração foi incorporada, separadamente, em ágar BDA, e vertida em placas de petri, seguido do disco de micélio do fungo, onde o diâmetro das colônias foi medido diariamente. Utilizou-se como controle negativo, ágar sem extrato e ágar com solução etanólica. O extrato etanólico de B. crassifolia apresentou atividade inibitória sobre os fungos estudados, onde as concentrações de 800 e 1600 µg 100 mL-1, inibiram 38% do crescimento micelial de F. solani; para S. sclerotiorum, a melhor concentração foi de 2400 µg 100 mL1, com 37,5% de redução de crescimento. Atribui-se o potencial antifúngico do extrato da casca da espécie aos compostos fenólicos e derivados de triterpenos.

Establishment of a tree shrew model of Fusarium keratitis / 中国实验动物学报

Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma.Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days.Fungal suspension was collected and the number of spores was adjusted to 1 × 1010 CFU /mL on the blood cell count plate.Forty healthy tree shrews were randomly divided into experimental group (n=30) and control group (n=10).In the experimental group, 50 μL of fungal spore suspension was injected into the cornea center with a 29G needle, and 50 μL saline was injected in the control group.The models were evaluated by anterior segment photography, in vivo confocal microscopy, histopathology, and corneal tissue culture.Results The fungal infiltration, the degree of edema of corneal epithelial and endothelial cells, and the number of mycelium were positively correlated with time.The number of infiltrating inflammatory cells, mainly, neutrophils, reached a peak on the 7th day after modeling.The mycelial growth was parallel to the stromal fibers.After the successful establishment of the model, the corneal tissue culture showed the growth of Fusarium solani.The successful rate of modeling was 86%.Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.

Changes of Th1/Th2 inflammatory factors in the aqueous humor of tree shrews with fusarium solani keratitis / 眼科新进展

Objective To observe the changes of Th1 / Th2 inflammatory factors in the aqueous humor of tree shrews with fusarium solani keratitis,as well as to explore the relationship between Thl / Th2 inflammatory factors and inflammatory response in fusarium solani keratitis.Methods Forty healthy tree shrews were randomly divided into experimental group(n =30) and control group (n =10).Fusarium solani was inoculated into sabina culture medium and cultured at 26 ℃ for 7 days,and then the fungal suspension was collected and the density of spores was adjusted to 10 × 109 CFU · mL-1.In the experimental group,50 μL fungal spore suspension was injected into the center of the cornea stroma,while the control group received the same amount of saline.Next the levels of cytokines including interleukin (IL)-1 β,IL-6,IL-4 and IL-10 were analyzed by flow cytometry on day 3,day 7,day 14 after successful modeling,and the changes in types of infiltrating cells were observed by histopathological examination.Results The expression level of IL-1 β and IL-6 (Th1 type cytokines) was the highest on day 7,and the difference was statistically significant at each time point when compared with the control group (all P ＜ 0.05).The expression level of IL-10 (Th2 type cytokines) was the highest on day 14,and the difference was statistically significant at each time point when compared with the control group (all P ＜ 0.05).The difference in IL-4 expression was statistically significant on day 7 (P ＜ 0.05).In addition,histopathological examination showed that the number of infiltration cell reached its peak on day 7,mainly neutrophils,and fungal hyphae were observed to be parallel to the matrix fibers at each time point.Conclusion The proinflammatory cytokines IL-1β and IL-6 and the anti-inflammatory cytokines IL-10 may play an important role in the molecular mechanism of inflammatory response of fusarium solani keratitis in tree shrews.

Chemotaxis response of Fusarium solani and Cylindrocarpon destructans on total ginsenoside / 中草药

Objective: To study the chemotaxis response of Fusarium solani and Cylindrocarpon destructans on total ginsenosides. Methods: Three chemotactic parameters (concentration, temperature, and pH) were determined by plate assay and spore germination method to research the chemotaxis response of two pathogens and their spores. Results: It showed that F. solani had strong chemotactic response at the low concentration of total ginsenosides, and the data of chemotactic mobile index (CMI) was 1.2948, SGR was 66%, chemotaxis growth rate (CGR) was 0.533, and (mycelial growth) MG was 0.5220 mg/mL. However, C. destructans had strong chemotactic response at the middle concentration of total ginsenosides, and the data of CMI was 1.2556, SGR was 63%, CGR was 0.465, and MG was 0.449 4 mg/mL. Conclusion: The low and middle concentration (0.2-20 mg/L) of total ginsenosides had the significant promoting effect on chemotaxis response of F. solani, and the chemotaxis response of C. destructans happened at different concentration of ginsenosides, and the SGR, MGR, and the amount of MG of these two pathogens had also been significantly improved, whereas the chemotaxis response effect decreases as the ginsenosides concentration increases.